Development of a Baltimore (MD) population of Calliphora vicina (Diptera: Calliphoridae) reared at several temperatures and estimations of developmental limits and thresholds.

Developmental data for necrophagous Diptera are frequently used in medico-legal investigations to estimate portions of the postmortem interval and interpret periods of insect activity. These applications require baseline developmental data for local populations from geographic locations of interest. For the widely distributed blow fly Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae), detailed developmental data does not exist for many locations in the mid-Atlantic region of the United States.

The effect of postmortem penetrative trauma on oviposition behavior of Calliphora vicina.

Flies in the family Calliphoridae are purported to demonstrate a strong attraction to and preferential colonization of wounds when present on human corpses or carrion. This well-circulated concept in Forensic Entomology is based on surprisingly few empirical studies that have examined the oviposition behavior of necrophagous Diptera toward wounds of vertebrate animals. In the present study, the oviposition behavior of Calliphora vicina toward piglets inflicted with postmortem sharp force trauma was examined during a 10-h test period under controlled laboratory conditions and in an outdoor urban environment.

Differential responses of adult Calliphora vicina to dry bloodstains on porous versus non-porous surface materials.

Necrophagous flies are presumed to feed on wet and dried blood at crime scenes, but no empirical information exists detailing fly interactions with dried bloodstains. In the present study, the foraging behavior of adult Calliphora vicina was characterized during interactions with dried bloodstains formed on a variety of porous, and non-porous materials that are commonly encountered in a household. Continuous digital recording and image analysis were used to monitor fly interactions with dried bloodstains and to determine mechanisms of stain modification.

Retention of Human Body Fluids in Adults of Calliphora vicina (Diptera: Calliphoridae).

Foraging by Calliphora vicina Robineau-Desvoidy often leads to a period of bubbling behavior, followed by either deposition of the regurgitate onto surfaces or reuptake of the bubble. Eventually, the partially or undigested food is passed in the excreta forming fecal or defecatory stains on surfaces in which deposition occurs. This study examined the digestive artifacts (i.

Characterization of Insect Stains Deposited by Calliphora vicina (Diptera: Calliphoridae) on Shirt Fabrics.

Despite the fact that necrophagous flies are known to alter bloodstains and create unique artifacts, no research has occurred to date that has examined the characteristics of insect stains on textiles or fabrics. This study represents the first effort to characterize artifacts produced by adult Calliphora vicina Robineau-Desvoidy deposited on a range of shirt fabrics that varied in type, color, orientation, and yarn tension. In general, artifact morphology on any type of fabric was distorted in comparison to those observed on smooth and/or nonporous surfaces in previous studies.

Detection of fly artifacts from four species of necrophagous flies on household materials using immunoassays.

An immunoassay was previously developed as a technique to improve methods for detection and analysis of fly artifacts found at crime scenes. The dot blot assay utilized a polyclonal antiserum (anti-md3) based on a unique digestive cathepsin D found in cyclorrhaphous Diptera. In this study, artifacts produced by adults of Calliphora vicina, Cynomya cadaverina, Sarcophaga bullata, and Protophormia terraenovae were examined using the immunoassay to determine if insect-derived stains could be distinguished from a range of human body fluid stains.

Immunoassay detection of fly artifacts produced by several species of necrophagous flies following feeding on human blood.

Foraging behavior of necrophagous flies commonly leads to distortion of human bloodstains and production of artifacts that confound reconstruction efforts at crime scenes. Currently there is no reliable method for detection of fly-derived stains or distinction of the artifacts from human bloodstains. To overcome these deficiencies, a confirmatory test was developed based on immunological detection of cathepsin D found in digestive fluids of and .

Distinction of Fly Artifacts from Human Blood using Immunodetection.

Insect stains produced by necrophagous flies are indistinguishable morphologically from human bloodstains. At present, no diagnostic tests exist to overcome this deficiency. As the first step toward developing a chemical test to recognize fly artifacts, polyclonal antisera were generated in rats against three distinct antigenic sequences of fly cathepsin D-like proteinase, an enzyme that is structurally distinct in cyclorrhaphous Diptera from other animals.

Morphological Features of Regurgitate and Defecatory Stains Deposited by Five Species of Necrophagous Flies are Influenced by Adult Diets and Body Size.

The morphological characteristics of artifacts from five species of necrophagous flies were examined following feeding on several types of diets. Four types of insect stains were produced by each species: regurgitate, defecatory, translocation, and tarsal tracks. Regurgitate was the most frequent type deposited (70.

First Record of the Larval Endoparasitoid Tachinaephagus zealandicus (Hymenoptera: Encyrtidae) From Fly Hosts Developing on Burnt Remains and in Pennsylvania.

Adults of the gregarious larval endoparasitoid Tachinaephagus zealandicus Ashmead (Hymenoptera: Encyrtidae) were collected from two species of carrion-inhabiting flies, Phormia regina Meigen (Diptera: Calliphoridae) and Hydrotaea aenescens Wiedemann (Muscidae), associated with the burnt remains of a domesticated pig, Sus scrofa L. (Artiodactyla: Suidae), during late summer in south central Pennsylvania. This represents the first reported occurrence of the wasp in the state and only the second in the mid-Atlantic region.

Spatial characterization of proteolytic enzyme activity in the foregut region of the adult necrophagous fly, Protophormia terraenovae.

The spatial distribution of proteolytic enzymes in the adult foregut of Protophormia terraenovae was studied in the context of protein digestion and regurgitation. Based on substrate specificity, pH optima, and use of specific protease inhibitors, all adults tested displayed enzyme activity in the foregut consistent with pepsin, trypsin and chymotrypsin. Chymotrypsin-like and trypsin-like enzyme activity were detected in all gut fluids and tissues tested, with chymotrypsin displaying the highest activity in saliva and salivary gland tissue, whereas maximal trypsin activity was evident in the crop.

Water balance characteristics of pupae developing in different size maggot masses from six species of forensically important flies.

The impact of maggot mass size on body water content, net transpiration rate, and dehydration tolerance of fly pupae was examined in six species of necrophagous flies. Species that spent more time on food as larvae, produced pupae with high body water contents. Dehydration tolerance limits of pupae were modest, matching the moisture-rich conditions of decaying carrion for larvae.

Oviposition restraint and developmental alterations in the ectoparasitic wasp, Nasonia vitripennis, when utilizing puparia resulting from different size maggot masses of Lucilia illustris, Protophormia terraenovae, and Sarcophaga bullata.

Adult females of the ectoparasitoid Nasonia vitripennis (Walker) are capable of distinguishing between hosts of different quality, and then correspondingly adjust clutch sizes and sex ratios of the offspring. In this study, we examined whether the size of the maggot mass, and presumably the developmental temperature, influenced the suitability of the resulting fly pupal and pharate adult stages as hosts for N. vitripennis.

Venom proteins from endoparasitoid wasps and their role in host-parasite interactions.

Endoparasitoids introduce a variety of factors into their host during oviposition to ensure successful parasitism. These include ovarian and venom fluids that may be accompanied by viruses and virus-like particles. An overwhelming number of venom components are enzymes with similarities to insect metabolic enzymes, suggesting their recruitment for expression in venom glands with modified functions.

Pathological and ultrastructural changes in cultured cells induced by venom from the ectoparasitic wasp Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae).

The ectoparasitic wasp Nasonia vitripennis produces a proteinaceous venom that induces death in fly hosts by non-paralytic mechanisms. Previous in vitro assays have suggested that the primary cause of cell and tissue death is oncosis, a non-programmed cell death (PCD) pathway characterized by cellular swelling and lysis. However, ultrastructural analyses of BTI-TN-5B1 cells exposed to LC(99) doses of wasp venom revealed cellular changes more consistent with apoptosis and/or non-apoptotic PCD than oncosis or necrosis: By 3h after incubation with venom, susceptible cells displayed indentations in the nuclear membranes, large nucleoli, and extensive vacuolization throughout the cytoplasm.

Changes in development and heat shock protein expression in two species of flies (Sarcophaga bullata [Diptera: Sarcophagidae] and Protophormia terraenovae [Diptera: Calliphoridae]) reared in different sized maggot masses.

Development of two species of necrophagous flies, Sarcophaga bullata Parker (Sarcophagidae) and Protophormia terraenovae (Robineau-Desvoidy) (Calliphoridae), was examined in different size maggot masses generated under laboratory conditions. Larvae from both species induced elevated mass temperatures dependent on the number of individuals per mass. The relationship was more evident for S.

Venom proteins of the parasitoid wasp Nasonia vitripennis: recent discovery of an untapped pharmacopee.

Adult females of Nasonia vitripennis inject a venomous mixture into its host flies prior to oviposition. Recently, the entire genome of this ectoparasitoid wasp was sequenced, enabling the identification of 79 venom proteins. The next challenge will be to unravel their specific functions, but based on homolog studies, some predictions already can be made.

The mode of action of venom from the endoparasitic wasp Pimpla hypochondriaca (hymenoptera: ichneumonidae) involves Ca+2-dependent cell death pathways.

The endoparasitoid Pimpla hypochondriaca injects venom during oviposition to condition its lepidopteran hosts. Venom is a complex mixture of proteins and polypeptides, many of which have been identified as enzymes, including phenoloxidase, endopeptidase, aminopeptidase, hydrolase, and angiotensin-converting enzyme. Constituents of the venom have been shown to possess cytolytic and paralytic activity, but the modes of action of factor(s) responsible for exerting such effects have not been deciphered.

Age and diet influence the composition of venom from the endoparasitic wasp Pimpla turionellae L. (Hymenoptera: Ichneumonidae).

Venom from the endoparasitic wasp Pimpla turionellae L. (Hymenoptera: Ichneumonidae) was found to contain a complex mixture of biogenic amines, noradrenalin, phospholipase B, and several proteins and peptides. The amount of noradrenalin and serotonin was found to be highest in venom from newly emerged wasps and decreased with age.

Characterization of phenoloxidase activity in venom from the ectoparasitoid Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae).

Crude venom isolated from the ectoparasitic wasp Nasonia vitripennis was found to possess phenoloxidase (PO) activity. Enzyme activity was detected by using a modified dot blot analysis approach in which venom samples were applied to nylon membranes and incubated with either L-DOPA or dopamine. Dot formation was most intense with dopamine as the substrate and no activators appeared to be necessary to evoke a melanization reaction.

In vivo and in vitro activity of venom from the endoparasitic wasp Pimpla turionellae (L.) (Hymenoptera: Ichneumonidae).

The biological activity of venom from Pimpla turionellae L. (Hymenoptera: Ichneumonidae) was examined in vivo toward larvae and pupae of Galleriae mellonella L. (Lepidoptera: Pyralidae), and in vitro toward bacterial and fungal cultures, as well as cultured insect cells.

Characterization and biochemical analyses of venom from the ectoparasitic wasp Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae).

During parasitism, the ectoparasitic wasp Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae) induces a developmental arrest in host pupae that is sustained until the fly is either consumed by developing larvae or the onset of death. Bioassays using fluids collected from the female reproductive system (calyx, alkaline gland, acid gland, and venom reservoir) indicated that the venom gland and venom reservoir are the sources of the arrestant and inducer(s) of death. Infrared spectroscopic analyses revealed that crude venom is acidic and composed of amines, peptides, and proteins, which apparently are not glycosylated.

Stress-induced accumulation of glycerol in the flesh fly, Sarcophaga bullata: evidence indicating anti-desiccant and cryoprotectant functions of this polyol and a role for the brain in coordinating the response.

Nondiapausing larvae of the flesh fly, Sarcophaga bullata, responded to several forms of short-term environmental stress (low temperature, anoxia and desiccation) by accumulating glycerol. Elevation of this polyol, regardless of the type of stress that induced accumulation, conferred cold resistance: larvae with high glycerol levels were 3-4 times more tolerant of a 2h exposure to -10 degrees C than unstressed larvae. Protection against low temperature injury, as well as dehydration, was also attained by injection of exogenous glycerol into third instar larvae.